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Molecular Analysis of Trypanosome Infection

锥虫感染的分子分析

基本信息

批准号：
7668688
负责人：
Fernando Villalta
金额：
$ 35.93万
依托单位：
MEHARRY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-01 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7668688
关键词：
Adhesions Affect Americas Amino Acids Animals Area Binding Blood Cardiac Cardiovascular system Cells Chagas Disease Chimera organism Chronic Computational Biology Computer Simulation Data Deletion Mutation Disease Event Extracellular Domain Extracellular Matrix Functional disorder Gastrointestinal tract structure Genes Goals Heart Diseases Human In Vitro Individual Infection Interruption Intervention Knock-out Knockout Mice Knowledge LOX gene Laminin Lectin Ligands Low Density Lipoprotein Receptor Mammalian Cell Mediating Modeling Molecular Molecular Analysis Morbidity - disease rate Mus Mutate Outcome Parasites Pathogenesis Pathology Peptides Preventive Process Proteins RNA Interference Receptor Cell Regulation Reporting Research Research Personnel Role Signal Pathway Signal Transduction Signal Transduction Pathway Structure Structure-Activity Relationship Surface Testing Therapeutic Intervention Time Tissues Transgenic Mice Transgenic Organisms Trypanosoma Trypanosoma cruzi Up-Regulation Work base heart function human morbidity in vivo innovation laminin-1 mortality mouse model novel novel strategies oxidized low density lipoprotein receptor receptor density receptor-mediated signaling response tool

项目摘要

DESCRIPTION (provided by applicant): Trypanosoma cruzi is a blood and tissue parasite that affects millions of individuals causing significant human morbidity and mortality. Basic understanding of the role of the critical surface molecules that participate in the first step of T. cruzi infection may provide novel targets for therapy. Our long-range goal is to understand the molecular mechanisms that allow T. cruzi to infect mammalian cells and cause disease, so that specific molecular intervention strategies can be developed against T. cruzi infection. The objective of this application is to identify which host cell receptors mediate T. cruzi binding to mammalian cells, when receptor-mediated signaling pathways contribute to infection. The hypothesis of this application is that T. cruzi gp83, a ligand molecule that the parasite uses to attach to mammalian cells, binds to the lectin-like oxidized low-density lipoprotein receptor (LOX-1) of mammalian host cells to mediate trypanosome attachment, thereby activating signaling events leading to initial infection and pathogenesis. We have formulated this hypothesis based on strong preliminary results, showing that interruption of LOX-1 gene by gene trapping or silencing LOX-1 gene by RNAi inhibits T. cruzi binding to cells and infection, whereas over-expression of LOX-1 in cells causes over-attachment of trypanosomes to cells and increased infection. Supporting also this hypothesis are our findings showing that gp83 specifically binds to LOX-1 and that we have identified critical regions on gp83 and LOX-1 that interact with each other to apparently mediate T. cruzi attachment leading to parasite entry. We will test our central hypothesis by pursuing the following specific aims: (1) to determine the structural-function relationships of the LOX-1-gp83 interaction; (2) to determine the LOX-1- dependent signaling pathways that mediate trypanosome gp83-dependent infection; and (3) to determine the in vivo role of the LOX-1 gene in the process of T. cruzi infection and pathogenesis using a novel LOX-1 knock out mouse model and a novel transgenic mouse model over-expressing LOX-1. Mutations, deletions and substitutions in critical regions of LOX-1 and gp83 will be performed to identify the motifs in each interacting molecule involved in T. cruzi binding to cells. Computational modeling of the trypanosome gp83 and the interacting peptide loop from the extracellular domain of LOX-1 will be determined. The mechanism underlying the novel gp83-LOX-1 signal transduction pathway leading to regulation of laminin ?-1 expression and infection will be elucidated using LOX-1(-/-) cells and LOX(+/+) cells. We will use our novel mouse models that over-express LOX-1 or that are null for LOX-1 expression to provide definitive in vivo tools to understand the other complementary mechanisms that contribute to trypanosome infection (in the LOX-1 null animals) and the mechanisms by which LOX-1 mediates infection (in the over-expressing animals).

描述（由申请方提供）：克氏锥虫是一种血液和组织寄生虫，影响数百万人，导致严重的人类发病率和死亡率。对参与T. cruzi感染可能提供新的治疗靶点。我们的长期目标是了解T。cruzi感染哺乳动物细胞并导致疾病，因此可以开发针对T.克氏感染本申请的目的是鉴定哪些宿主细胞受体介导T.当受体介导的信号传导途径导致感染时，cruzi与哺乳动物细胞结合。本申请的假设是T. cruzi gp 83是寄生虫用于附着于哺乳动物细胞的配体分子，其与哺乳动物宿主细胞的凝集素样氧化低密度脂蛋白受体（LOX-1）结合以介导锥虫附着，从而激活导致初始感染和发病的信号传导事件。我们基于强有力的初步结果阐述了这一假设，表明通过基因诱捕阻断LOX-1基因或通过RNAi沉默LOX-1基因抑制T.细胞中LOX-1的过度表达导致锥虫与细胞的过度附着和感染增加。我们的发现也支持这一假设，表明gp 83特异性结合LOX-1，并且我们已经鉴定了gp 83和LOX-1上相互作用以明显介导T的关键区域。克氏附着导致寄生虫进入。我们将通过以下具体目标来验证我们的中心假设：（1）确定LOX-1-gp 83相互作用的结构-功能关系;（2）确定介导锥虫gp 83依赖性感染的LOX-1依赖性信号通路;（3）确定LOX-1基因在T.使用新的LOX-1敲除小鼠模型和新的过表达LOX-1的转基因小鼠模型研究克氏克氏病感染和发病机制。LOX-1和gp 83关键区域的突变、缺失和取代将被执行以鉴定参与T. cruzi与细胞结合。将确定锥虫gp 83和LOX-1胞外结构域的相互作用肽环的计算建模。新的gp 83-LOX-1信号转导通路调节层粘连蛋白？1的表达和感染将使用LOX-1（-/-）细胞和LOX（+/+）细胞阐明。我们将使用我们的过表达LOX-1或LOX-1表达无效的新型小鼠模型来提供确定的体内工具，以了解有助于锥虫感染的其他互补机制（在LOX-1无效动物中）和LOX-1介导感染的机制（在过表达动物中）。